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pET28a-HA-CPDSalI
(Plasmid #38254)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 38254 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET28a
  • Backbone manufacturer
    Novagen
  • Backbone size (bp) 6029
  • Modifications to backbone
    HA epitope tag and Vibrio cholerae MARTX toxin (rtxA, vc1451) cysteine protease domain (CPD) (amino acids 3440-3650) cloned into SalI and XhoI sites in backbone to create fusions with target protein of interest
  • Vector type
    Bacterial Expression
  • Promoter T7
  • Tags / Fusion Proteins
    • HA (C terminal on backbone)
    • Vibrio cholerae MARTX toxin cysteine protease domain (CPD) (C terminal on backbone)
    • 6xHis (C terminal on backbone)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ sequencing primer T7 promoter
  • 3′ sequencing primer T7 terminator
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

For protein purification, target protein of interest should be cloned into NcoI and SalI sites to create HA-tagged fusion with Vibrio cholerae MARTX toxin cysteine protease domain (CPD). HA epitope tag will remain on target protein after cleavage.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET28a-HA-CPDSalI was a gift from Matthew Bogyo & Aimee Shen (Addgene plasmid # 38254 ; http://n2t.net/addgene:38254 ; RRID:Addgene_38254)
  • For your References section:

    Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag. Shen A, Lupardus PJ, Morell M, Ponder EL, Sadaghiani AM, Garcia KC, Bogyo M. PLoS One. 2009 Dec 2;4(12):e8119. 10.1371/journal.pone.0008119 PubMed 19956581